Mercury-Modulated Immune Responses in Arctic Barnacle Goslings (Branta leucopsis) upon a Viral-Like Immune Challenge.

Historical mining activities in Svalbard (79°N/12°E) have caused local mercury (Hg) contamination. To address the potential immunomodulatory effects of environmental Hg on Arctic organisms, we collected newborn barnacle goslings (Branta leucopsis) and herded them in either a control or mining site, differing in Hg levels. An additional group at the mining site was exposed to extra inorganic Hg(II) via supplementary feed. Hepatic total Hg concentrations differed significantly between the control (0.011 ± 0.002 mg/kg dw), mine (0.043 ± 0.011 mg/kg dw), and supplementary feed (0.713 ± 0.137 mg/kg dw) gosling groups (average ± standard deviation). Upon immune challenge with double-stranded RNA (dsRNA) injection, endpoints for immune responses and oxidative stress were measured after 24 h. Our results indicated that Hg exposure modulated the immune responses in Arctic barnacle goslings upon a viral-like immune challenge. Increased exposure to both environmental as well as supplemental Hg reduced the level of natural antibodies, suggesting impaired humoral immunity. Hg exposure upregulated the expression of proinflammatory genes in the spleen, including inducible nitric oxide synthase (iNOS) and interleukin 18 (IL18), suggesting Hg-induced inflammatory effects. Exposure to Hg also oxidized glutathione (GSH) to glutathione disulfide (GSSG); however, goslings were capable of maintaining the redox balance by de novo synthesis of GSH. These adverse effects on the immune responses indicated that even exposure to low, environmentally relevant levels of Hg might affect immune competence at the individual level and might even increase the susceptibility of the population to infections.

Modulatory Effects of Mercury (II) Chloride (HgCl2 ) on Chicken Macrophage and B-Lymphocyte Cell Lines with Viral-Like Challenges In Vitro.

Mercury (Hg) is a toxic trace metal ubiquitously distributed in the environment. Inorganic mercury (as HgCl2 ) can cause immunotoxicity in birds, but the mechanisms of action are still not fully resolved, especially with respect to responses to viral infections. To investigate the potential immunomodulatory effects of Hg2+ on specific cell types of the avian immune system, chicken macrophage (HD-11) and B-lymphocyte (DT40) cell lines were applied as in vitro models for the innate and adaptive immune systems, respectively. The cells were stimulated with synthetic double-stranded RNA, which can be recognized by toll-like receptor-3 to mimic a viral infection. The Hg2+ showed concentration-dependent cytotoxicity in both cell lines, with similar median effect concentrations at 30 µM. The cytotoxicity of Hg2+ was closely related to glutathione (GSH) depletion and reactive oxygen species induction, whereas the de novo synthesis of GSH acted as a primary protective strategy. Nitric oxide produced by activated macrophages was strongly inhibited by Hg2+ , and was also influenced by cellular GSH levels. Cell proliferation, gene expression of microRNA-155, and cellular IgM levels in B cells were decreased at noncytotoxic Hg2+ concentrations. The secretion of antiviral interferon-α was induced by Hg2+ in both cell lines. Overall, our results suggest that Hg2+ exposure can cause immunomodulatory effects in birds by disrupting immune cell proliferation and cytokine production, and might result in disorders of the avian immune system. Environ Toxicol Chem 2021;40:2813-2824. © 2021 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Modulatory Effects of Pb2+ on Virally Challenged Chicken Macrophage (HD-11) and B-Lymphocyte (DT40) Cell Lines In Vitro.

Elevated levels of lead have been found in waterfowl, due to human activities. Lead may cause immunomodulatory effects, but the mechanisms are largely unknown, especially after viral challenges. To characterize avian immunomodulatory hazards of lead (Pb)2+ , we used chicken macrophage (HD-11) and B-lymphocyte (DT40) cell lines, as in vitro models for the innate and adaptive immune systems, respectively. The cells were activated via toll-like receptor-3 by polyinosinic-polycytidylic acid sodium salt (poly I:C), mimicking viral infections. Our results indicate that Pb2+ is cytotoxic to both cell lines, macrophages being more sensitive. De novo synthesis of glutathione plays an important role in protecting macrophages from Pb2+ intoxication, which might also be closely involved in the induction of nitric oxide after Pb2+ exposure. Stimulatory effects on cell proliferation were noticed at noncytotoxic Pb2+ concentrations as well. Exposure to Pb2+ could also affect the inflammatory status by inhibiting the pro-inflammatory interferon (IFN)-γ while promoting the production of anti-inflammatory type I IFNs in both macrophages and B-cells, and increasing intracellular IgM levels in B-cells. These results suggest that the immunomodulatory effects of Pb2+ in birds are probably closely associated with disruption of immune cell proliferation and cytokine production, potentially causing disorders of the avian immune system. Environ Toxicol Chem 2020;39:1060-1070. © 2020 SETAC.

Cell-specific immune-modulation of cadmium on murine macrophages and mast cell lines in vitro.

Toxic trace metals are widespread contaminants that are potentially immunotoxic even at environmentally low exposure levels. They can modulate the immunity to infections, e.g., in wildlife species living in contaminated areas. The diverse immune cell types can be differentially affected by the exposure leading to the modulation of specific protective mechanisms. Macrophages and mast cells, part of the innate immune system, trigger immune responses and perform particular effector functions. The present study compared toxicological and functional effects of cadmium in two models of murine macrophages (RAW264.7 and NR8383 cell lines) and two models of murine mast cells (MC/9 and RBL-2H3 cell lines). Cadmium was selected as a model compound because its known potential to induce reactive oxygen species and its relevance as an environmental contaminant. Mechanisms of toxicity, such as redox imbalance and apoptosis induction were measured in stationary cells, while functional outcome effects were measured in activated cells. Cadmium-depleted glutathione antioxidant in all four cell lines tested although reactive oxygen species was not significantly increased. Mast cells had full dose-response depletion of glutathione below cytotoxic levels while in macrophages the depletion was not complete. Functional endpoints tumour necrosis factor-alpha and nitrite production in lipopolysaccharide-activated macrophages were increased by cadmium exposure. In contrast, mast cell lipopolysaccharide-induced tumour necrosis factor-alpha and IgE-mediated histamine release were reduced by cadmium. These data indicate potentially differential effects of cadmium among murine innate immune cell types, where mast cells would be more susceptible to oxidative stress and their function might be at a higher risk to be modulated compared to macrophages.

Injectable chitosan/ß-glycerophosphate hydrogels with sustained release of BMP-7 and ornidazole in periodontal wound healing of class III furcation defects.

The aim of this study was to determine the effect of bone morphogenetic protein-7 (BMP-7) and ornidazole (ORN) loaded Chitosan/ß-glycerophosphate (CS/ß-GP) thermosensitive hydrogels on periodontal regeneration. CS/ß-GP hydrogels with and without BMP-7 and ORN were compared with respect to physicochemical properties, release kinetics, and antimicrobial activity in vitro, and periodontal regeneration properties in class III furcation defects in beagles via radiography, histology including immunohistochemical staining of osteoblasts and osteoclasts, and histometric analysis. CS/ß-GP hydrogels with and without BMP-7 and ORN had comparable physicochemical properties and gelation kinetics. Release kinetics showed that the hydrogels were capable of stable and sustained release of BMP-7 and ORN. The hydrogels loaded with ORN exhibited obvious antimicrobial activity against P. gingivalis. Histometric analysis quantitatively showed significantly more new bone and cementum, and less connective tissue in defects implanted with BMP-7 loaded hydrogels compared with hydrogels without BMP-7. The number of osteoclasts reduced significantly in the CS/BMP-7/ORN and CS/BMP-7 groups, while the number of osteoblasts increased significantly in these groups. Our findings showed that BMP-7 and ORN conferred additional advantages to the CS/ß-GP hydrogel in periodontal regeneration and suggest potential consideration of this approach for periodontal therapy.

Chitosan composite scaffold combined with bone marrow-derived mesenchymal stem cells for bone regeneration: in vitro and in vivo evaluation.

The study aimed to develop a chitosan (CS)-based scaffold for repairing calvarial bone defects. We fabricated composite scaffolds made of CS and bovine-derived xenograft (BDX), characterized their physicochemical properties including pore size and porosity, absorption, degradation, and compressive strength, compared their efficacy to support in vitro proliferation and differentiation of human jaw bone marrow-derived mesenchymal stem cells (hJBMMSCs), and evaluated their bone regeneration capacity in critical-size rat calvarial defects. The CS/BDX (mass ratio of 40:60) composite scaffold with porosity of 46.23% and pore size of 98.23 µm exhibited significantly enhanced compressive strength than the CS scaffold (59.33 ± 4.29 vs. 18.82 ± 2.49 Kpa). The CS/BDX (40:60) scaffold induced better cell attachment and promoted more osteogenic differentiation of hJBMMSCs than the CS scaffold. The CS/BDX (40:60) scaffold seeded with hJBMMSCs was the most effective in supporting new bone formation, as evidenced by better histomorphometry results, larger new bone area, and more obvious mature lamellar bone formation compared to other groups in rat calvarial defects 8 weeks after implantation. These results suggest that CS/BDX composite scaffold combining with hJBMMSCs has the potential for bone defect regeneration.